Autoimmunity is an important cause of disease affecting 3-5% of the human population. In particular, 58 different neurological diseases with autoimmune etiology have been described. Although some factors causing these diseases have been identified, the etiology of most of autoimmune diseases remains obscure. Nevertheless, the antibody response to self antigens is considered indicative of an anti-self response.

Only in a few cases the antigen responsible for the rupture of the self-tolerance has been characterized. This is particularly important for the diagnosis, since when an autoantigen is identified as specific marker, diagnostic tests based on the purified antigen are used. In all other cases, an immunofluorescence test is performed on histological sections challenged with the patient’s serum but only limited information on the antigen is obtained.

Objectives of this UO is to develop approaches to describe the complexity of the humoral response in autoimmune diseases. The antigens identified are produced as recombinant proteins to develop new diagnostic assays. In addition we are creating an image data bank to characterize the different staining pattern produced by sera from neurological patients using semiautomated immunohistochemical analysis of the reactivity of sera on rat brain sections.

2.Neurotrophic Factors as biomarkers and drug-targets for neurological diseases

Brain-Derived Neurotrophic Factor (BDNF) is a growth and trophic factor that belongs to the family of related secretory proteins called neurotrophins. BDNF plays multifaceted and in part opposed functions in both development and maintenance of the nervous system. Indeed, BDNF promotes both cell survival and cell death, neuronal maturation including neurites and dendritic spines outgrowth, and has a prominent role in various forms of synaptic plasticity such as long-term potentiation and long-term depression (Casaccia-Bonnefil et al., 1999; Huang and Reichardt, 2001; Lu et al., 2005). BDNF has a complex biology as it is composed of 22 transcripts in rodents (34 in humans) each of which contain one of the 11 different exons of BDNF gene encoding the 5’untranslated and a common downstream eaxon containing the coding region and the 3’untranslated regions (UTRs) (Aid et al., 2007).

The need and importance is increasingly felt for the identification of specific compounds able to modulate BDNF, which can be used as drugs for the treatment of neurological and neuropsychiatric diseases or for the treatment of deleterious effects caused to the nervous system by abuse of illegal or legal drugs. It is therefore object of the present invention the development of a method of screening for BDNF translation modulators which allow the measurement of BDNF protein production. In particular it would be highly desirable to identify a screening method which would allow to determine the expression of all possible BDNF variants, and allow, at the same time, to obtain information on the final amounts of the BDNF protein produced in response to a specific drug.

TECNOLOGIE IN POSSESSO DELL'U. O.

Phage display

Bacterial two-hybrid system

Recombinant proteins production

Two-D gel proteomic analysis

Medium-scale cell survival assay

Medium-scale bioluminescent assay

Medium-scale semi-automated immunohystochemical analysis

Confocal microscopy

Electron microscopy

STRUMENTAZIONE

Denominazione

Struttura ove la strumentazione è allocata

Responsabile della strumentazione

Confocal microscope

Dip. Scienze della Vita

E. Tongiorgi

Nikon E800 light microscope

Dip. Scienze della Vita

E. Tongiorgi

Nikon LE300 inverted light microscope with micromanipulators and chamber for live cell analysis